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  • Open Daily: 10am - 10pm
    Alley-side Pickup: 10am - 7pm

    3038 Hennepin Ave Minneapolis, MN
    612-822-4611

Open Daily: 10am - 10pm | Alley-side Pickup: 10am - 7pm
3038 Hennepin Ave Minneapolis, MN
612-822-4611
Characterization of Clostridium Perfringens Beta2 Toxin.

Characterization of Clostridium Perfringens Beta2 Toxin.

Gurjar, Abhijit

Paperback

Biology

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ISBN10: 1243539879
ISBN13: 9781243539878
Publisher: Proquest Umi Dissertation Pub
Pages: 162
Weight: 0.74
Height: 0.42 Width: 7.99 Depth: 10.00
Language: English
In recent years, a variant of the Clostridium perfringens beta toxin known as beta2 has been widely reported as one of the causative agents in C. perfringens related enteric diseases. The clinical symptoms of beta2 toxin induced enterotoxemias are characterized by watery sporadic diarrhea, edema of affected tissues and loss of body weight. The role of beta2 toxin in the causation of enterotoxemias and or haemorrhagic/necrotic enteritis in humans, domestic animals, birds and aquatic species has generated considerable interest in the field of human and veterinary medicine. Clostridium perfringens beta2 has been reported as an accessory toxin and may act in synergy with other major toxins of the C. perfringens in the production of necrotic and hemorrhagic enteritis. The beta2 toxin may play a role in hindering the process of nutrient absorption in the intestine and thereby debilitating affected animals. It is strongly speculated that a sudden change in the gut Microbiolflora or the intestinal physiological equilibrium may lead to clostridial disease outbreak. Studies that report on C. perfringens beta2 producing isolates from healthy animals suggest that beta2 toxin as a potential health hazard. Very few studies have reported the biophysical and molecular biological properties of the beta2 toxin in detail. It has been hypothesized that the mechanism of the beta2 toxin induced enteritis could include direct effects on transport of ions in the intestinal epithelial cells through membrane binding or pore formation. It is felt that experiments that focus on studying the cytotoxicity of beta2 toxin could lead to better understanding of beta2 toxin in context of C. perfringens related enterotoxemias. Based on these reported findings, elucidation of the molecular and biological properties of beta2 toxin would be of significant value to researchers towards developing a targeted epitope vaccine. Clostridium perfringens are toxinotyped into types A-E by conventional bacteriology and PCR based confirmation techniques. Owing to the widespread distribution of C. perfringens toxinotypes and its epidemiological significance a highly sensitive diagnostic tool that replaces the conventional PCR is required. In food animal practice, active immunoprophylaxis against C. perfringens is achieved by use of commercial toxoid vaccines against types B, C and D. There are no published reports on vaccines conferring immunity against beta2 toxin. The primary objectives of this dissertation are (1) develop a molecular based toxin typing technique for C. perfringens which has practical application in a diagnostic laboratory and aid in conducting molecular epidemiological studies, (2) Elucidate the structure and function of beta2 toxin, (3) Evaluate in vitro the cytotoxic properties of beta2 toxin, and (4) Evaluate the role of beta2 synthetic lipopeptide vaccine in a challenge study in the mouse model. The results of our study revealed a high prevalence and distribution of C. perfringens beta2 toxin encoding strains in feces of lactating cattle. Among the five toxinotypes, toxinotype A was widely distributed among seven dairy herds screened by multiplex Real time PCR assay. We successfully used a GST fusion system for expression of recombinant beta2 toxin. The purified recombinant toxin was shown to be cytotoxic to CaCo2 cell line at concentration of more than 10 mug/ml. the toxin treated cells exhibited cytopathic effects such as cell rounding, membrane blebbing and leakage of cytoplasm. These results could suggest that the C. perfringens beta2 toxin could act as one of the pore forming toxins. A monoclonal antibody raised against beta2 toxin completely...

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